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Utilization of multiple real-time PCR assays for the diagnosis of Bordetella spp. in clinical specimens

Kathleen M Tatti1, Maria Lucia Tondella

  • 1Division of Bacterial Diseases, Centers for Disease Control and Prevention, National Center for Immunization and Respiratory Diseases, Meningitis and Vaccine Preventable Diseases Branch, Atlanta, GA, USA. ket2@cdc.gov

Methods in Molecular Biology (Clifton, N.J.)
|October 30, 2012
PubMed

Insights

Bordetella pertussis, a cause of upper respiratory infections, can now be accurately diagnosed faster using real-time PCR assays. This study details specimen collection, DNA extraction, and PCR methods for detecting Bordetella species.

Area of Science:

  • Medical Microbiology
  • Infectious Diseases
  • Molecular Diagnostics

Background:

  • Pertussis, caused by Bordetella pertussis, is a significant respiratory infection affecting all age groups.
  • Accurate and timely diagnosis of pertussis can be challenging, impacting treatment and public health control.
  • The advent of molecular diagnostic techniques has improved the detection of infectious agents.

Purpose of the Study:

  • To describe a validated method for the detection and identification of Bordetella species using real-time PCR.
  • To outline the procedures for nasopharyngeal specimen collection and DNA extraction suitable for PCR analysis.
  • To facilitate earlier and more accurate clinical diagnosis of pertussis.

Main Methods:

  • Collection of nasopharyngeal specimens from patients with suspected pertussis.
  • Extraction of bacterial DNA from collected specimens.
  • Utilization of real-time polymerase chain reaction (PCR) assays for the sensitive detection of Bordetella DNA.

Main Results:

  • The described real-time PCR method allows for the specific detection of Bordetella species.
  • The protocol enables rapid identification of the causative agent of pertussis.
  • Implementation of this assay enhances diagnostic capabilities in clinical laboratories.

Conclusions:

  • Real-time PCR assays provide a rapid and accurate method for diagnosing Bordetella pertussis infections.
  • Standardized methods for specimen handling and DNA extraction are crucial for reliable molecular diagnostics.
  • This approach aids in the timely management of pertussis and supports public health surveillance.

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